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Sang Jin Kim, Jung Hyun Park, Un Chul Park, Myung Soo Cho, Haksup Lee, Seung-Yup Ku, Hyeong Gon Yu; Generation of Retinal Pigment Epithelial Cells from Human Embryonic Stem Cell-derived Spherical Neural Masses. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2227.
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© ARVO (1962-2015); The Authors (2016-present)
Using spherical neural masses (SNMs), pure cell masses of embryonic stem cells derived neural precursors, is efficient and prompt method of generating functional neurons. For retinal cell replacement therapy, we regenerated functional retinal pigment epithelial (RPE) cells from human embryonic stem cells derived SNMs. We investigated morphological, molecular, and functional characteristics of RPEs from hESC-derived SNMs.
RPEs were differentiated from human embryonic stem cells (SNUhES3) derived SNMs by neuroectodermal induction and mechanical purification. Morphologic evaluation was done with light microscopy and transmission electron microscopy. Expression of molecular markers was analyzed by RT-PCR and immunocytochemistry. After incubation of the cells with FITC-conjugated latex beads and bovine photoreceptor outer segment, confocal microscopic exam were done to evaluate phagocytosis.
Most of the differentiated cells from hESC derived SNMs showed typical RPE morphologies such as polygonal shaped epithelial monolayer with pigmentation. Transmission electron microscopic exam revealed apical microvilli, pigment granules, and tight junctions. RPE cells derived from SNMs also expressed cellular and molecular markers, Pax6, Mitf, ZO-1, RPE65, and bestrophin. Regenerated RPE function was confirmed by analysis of phagocytosis using latex bead and isolated bovine photoreceptor outer segment.
RPEs were efficiently generated from hESC derived SNMs. The cells showed specific morphology, expression of molecular markers, and phagocytosis of photoreceptor outer segment.
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